Derivation of germ cells using an adherent feeder-free culture system

This protocol describes a method for derivation of germ-like cells from human pluripotent stem cells. In the first step of the protocol, stem cells are virally transduced with vectors encoding for the green fluorescence protein gene under the control of the VASA promoter, which is a selective germ cell marker. This enables isolation of a subpopulation of VASA-expressing germ-like cells in the last step of the protocol. Alternatively, it is possible to different pluripotent stem cells in the differentiation medium for 7-14 days, without the viral transduction and cell sorting steps.